Difference between revisions of "Team:Reading/Experiments"

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<p>Note: This page is an account of all protocols used by the team this year. The references at the bottom of the page provide the sources for the protocols, however we have modified several of them to our needs.</p>
 
<p>Note: This page is an account of all protocols used by the team this year. The references at the bottom of the page provide the sources for the protocols, however we have modified several of them to our needs.</p>
  
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<p><h4>Growth of <i>Synechocystis sp. PCC 6803</i></h4>
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<p><h5>Growth of <i>Synechocystis sp. PCC 6803</i></h5>
 
<ol>
 
<ol>
 
<li>Thaw stock of 6803 on ice</li>
 
<li>Thaw stock of 6803 on ice</li>
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</ol></p>
 
</ol></p>
 
   
 
   
 +
<p><h5>Taking optical density of culture to measure growth</h5>
 +
We measured the OD<sub>750</sub> to match that used in research papers on <i>Synechocystis</i><sup>4</sup>.
 +
<ol>
 +
<li>Set spectrophotometer to measure at OD<sub>750</sub></li>
 +
<li>Blank with 1ml of BG-11</li>
 +
<li>Take OD<sub>750</sub> of 1ml of culture</li>
 +
<li>If OD<sub>750</sub> over 1 a.u, dilute 250µl of culture in 750µl of BG-11 (1/4 dilution)</li>
 +
</ol>
 +
To calculate cell density, we used the equation,
 +
1 a.u = 1.6x10<sup>8</sup> cells per ml of culture.
 +
  
 
<h5>What should this page contain?</h5>
 
<h5>What should this page contain?</h5>

Revision as of 16:51, 18 July 2015



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Protocols

Note: This page is an account of all protocols used by the team this year. The references at the bottom of the page provide the sources for the protocols, however we have modified several of them to our needs.

Growth of Synechocystis sp. PCC 6803
  1. Thaw stock of 6803 on ice
  2. 100ml of BG-11 medium added to sterile conical flask aseptically
  3. Add 10µl or 5 loopfuls of 6803 to flask
  4. Incubate at ~30°C and on shaker at ~50rpm, under constant illumination
  5. Check growth each day by measuring OD750

Taking optical density of culture to measure growth
We measured the OD750 to match that used in research papers on Synechocystis4.
  1. Set spectrophotometer to measure at OD750
  2. Blank with 1ml of BG-11
  3. Take OD750 of 1ml of culture
  4. If OD750 over 1 a.u, dilute 250µl of culture in 750µl of BG-11 (1/4 dilution)
To calculate cell density, we used the equation, 1 a.u = 1.6x108 cells per ml of culture.
What should this page contain?
  • Protocols
  • Experiments
  • Documentation of the development of your project

References